Visualization of dynamic interlinking networks which respond and adapt to the constantly changing environment would be highly beneficial in developing new composite materials and active/responsive materials. Here, optically and structurally stabilized plasmo‐bio interlinking networks (PBINs) free from photobleaching for high resolution, long term visualization are reported. Necessary for structural and optical stability, a new stability algorithm to comprehensively quantify stability and detect minute instability undetectable by traditional methods is introduced. Biocompatible plasmonic gold nanorods (Bio‐AuNRs) are synthesized for high resolution, long term imaging by utilizing bromide‐free alternatives to achieve CTA+ free. Systematic physical, chemical, and biological characterizations reveal the structural and optical stability of Bio‐AuNRs required for constructing PBIN. Lastly, with actin as a model of interlinking networks of the cytoskeleton, optically and structurally stable PBIN (100% CTA+ free, 97% crosslinking rate) in applications as active/responsive materials, are demonstrated.

CRISPR-Cas systems (clustered regularly interspaced short palindromic repeats) have shown great potential as efficient gene editing tools in disease therapeutics. Although numerous CRISPR-Cas systems have been developed, detailed mechanisms of target recognition and DNA cleavage are still unclear. In this work, we dynamically observe the entire process of conjugation, target recognition and DNA cleavage by single particle spectroscopy of CRISPR-Cas systems on single particle surfaces (gold) with the unique advantage of extended time periods. We show the CRISPR-Cas system, comprised of Cas endonuclease and single guide RNA, is stable and functional on single particle surfaces. Owing to the photostability of single particle surfaces, we directly observe in real time the entire dynamic process of conjugation, target recognition and DNA cleavage without photobleaching. We find heterogeneity in target recognition and DNA cleavage processes in which individual spectra vary significantly from one another as well as from the ensemble. We believe an in depth understanding of heterogeneities in CRISPR-Cas systems can overcome potential barriers in precision medicine and personalized disease therapeutics.

The crowded intracellular environment of biomolecules, including organelles, solutes, proteins, and membranes, presents distinct biomolecular dynamics crucial for the functions of biomolecules within living cells. However, background suppression is critical to uncover nanoscale dynamics in living cells. Herein, a new method for enabling rapid, nanoscale background elimination of cellular metallic nanoprobes is presented. By employing integrated nanoscopic correction (iNC) designed to eliminate depolarization effects, which compromise background elimination, a real‐time algorithm to subtract and increment orthogonal pairs of polarizations for real‐time nanoscale background elimination is introduced. The ability to analyze orthogonal pairs at high speed over the entire polarization range is currently difficult to achieve using conventional methods. By processing orthogonal pairs in real time, this method minimizes movement artifacts during the background elimination process. Nanometer spatial stability which enables two orders of magnitude increase in signal‐to‐noise ratio of cellular metallic nanoprobes is shown. Nanoscale background elimination aiding the ability to accurately track biomolecules and their dynamics in living cells is anticipated.